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IRCCS San Raffaele Scientific Institute

5 ARTICLES PUBLISHED IN JoVE

Cancer Research

Immunoglobulin Gene Sequence Analysis In Chronic Lymphocytic Leukemia: From Patient Material To Sequence Interpretation

Andreas Agathangelidis^*1, Lesley Ann Sutton^*2,3, Anastasia Hadzidimitriou¹, Cristina Tresoldi⁴, Anton W. Langerak⁵, Chrysoula Belessi⁶, Frederic Davi⁷, Richard Rosenquist^2,3, Kostas Stamatopoulos^1,2, Paolo Ghia⁸

¹Institute of Applied Biosciences, Centre for Research and Technology Hellas, ²Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, ³Department of Molecular Medicine and Surgery, Karolinska Institutet, ⁴Division of Immunology, Transplantation and Infectious, IRCCS San Raffaele Scientific Institute, ⁵Department of Immunology, Laboratory for Medical Immunology, Erasmus University Medical Center, ⁶Hematology Department, Nikea General Hospital, ⁷Assistance publique - Hôpitaux de Paris (AP-HP), Hopital Pitié-Salpêtrière, Department of Hematology, and UPMC University Paris 06, UMRS 1138, ⁸Division of Experimental Oncology, IRCCS Istituto Scientifico San Raffaele and Università Vita-Salute San Raffaele

Herein, we present a protocol that details the technical aspects and essential requirements to ensure robust IG gene sequence analysis in patients with chronic lymphocytic leukemia (CLL), based on the accumulated experience of the European Research initiative on CLL (ERIC).

Immunology and Infection

Purification and Expansion of Mouse Invariant Natural Killer T Cells for in vitro and in vivo Studies

Gloria Delfanti¹, Alessandra Perini^*1,2, Elisa Zappa^*1,2, Maya Fedeli^1,2

¹Experimental Immunology Unit, Division of Immunology, Transplantation and Infectious Diseases, IRCCS San Raffaele Scientific Institute, ²Vita-Salute San Raffaele University

We describe a rapid and robust protocol to enrich invariant natural killer T (iNKT) cells from mouse spleen and expand them in vitro to suitable numbers for in vitro and in vivo studies.

Medicine

Fertility Sparing Procedure using Carbon Dioxide Fiber Laser Vaporization of Ovarian Endometrioma

Massimo Candiani¹, Jessica Ottolina¹, Iacopo Tandoi¹, Ludovica Bartiromo¹, Matteo Schimberni¹, Roberta Villanacci¹, Stefano Ferrari¹

¹Gynecological Surgery and Endometriosis Departmental Unit, San Raffaele Scientific Institute

In this protocol, CO₂fiber laser technique is demonstrated for the surgical treatment of ovarian endometriosis, which represents a viable alternative in terms of fertility preservation with the major advantage of not being dependent on the surgeon's skills and personal experience.

Medicine

Laparoscopic Oocyte Retrieval and Cryopreservation during Vaginoplasty for Treatment of Mayer-Rokitansky-Kuster-Hauser Syndrome

Valeria Stella Vanni^*1,2, Alessandra Alteri^*1,2, Lucia De Santis^1,2, Greta Chiara Cermisoni¹, Elisa Rabellotti¹, Diana Delprato¹, Marta Parma¹, Enrico Papaleo¹, Luigi Fedele³, Massimo Candiani^1,2

¹Obstetrics and Gynaecology Unit, IRCCS San Raffaele Scientific Institute, ²Vita-Salute San Raffaele University, ³Department of Obstetrics and Gynecology, Fondazione IRCCS Ca' Granda, Ospedale Maggiore Policlinico

A dedicated team can offer Mayer-Rokitansky-Kuster-Hauser patients the option to perform controlled ovarian stimulation and oocyte cryopreservation at the time of laparoscopic vaginoplasty.

Bioengineering

In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing

Alessandro Migliara¹, Martino Alfredo Cappelluti¹, Francesca Giannese², Sara Valsoni¹, Alberto Coglot¹, Ivan Merelli^1,3, Davide Cittaro², Angelo Lombardo^1,4

¹San Raffaele Telethon Institute for Gene Therapy (SR-Tiget), IRCCS San Raffaele Scientific Institute, ²Center for Omics Sciences, IRCCS San Raffaele Institute, ³Institute for Biomedical Technologies, National Research Council, ⁴Vita-Salute San Raffaele University

Here, we present a protocol for the in vitro selection of engineered transcriptional repressors (ETRs) with high, long-term, stable, on-target silencing efficiency and low genome-wide, off-target activity. This workflow allows for reducing an initial, complex repertoire of candidate ETRs to a short list, suitable for further evaluation in therapeutically relevant settings.